In vitro culture systems for hepatitis B and delta viruses

  • Camille Sureau
Part of the Archives of Virology Supplementum book series (ARCHIVES SUPPL, volume 8)


The development of tissue culture technology has led to invaluable information in many fields of modern virology. Until recently, the lack of an in vitro culture system for the hepatitis B virus (HBV) was a considerable impediment to the study of its life cycle at the cellular and molecular levels. However, it did not prevent its isolation and molecular cloning. Such has been the case also for the hepatitis delta virus (HDV), the genome of which was cloned and sequenced before its replication could be observed in cultured cells. In recent years, tissue culture systems for HBV and HDV have been developed progressively by the identification of permissive, established cell lines for production of virions and susceptible primary hepatocyte cultures for infection assays. I will briefly review here the recent experiments that have contributed to replicate HBV and HDV in cell culture systems.


Huh7 Cell Human Hepatoma Cell Line Hepatitis Delta Virus Duck Hepatitis Dane Particle 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Acs G, Sells MA, Purcell RH, Price P, Engle R, Shapiro M, Poper H (1987) Hepatitis B virus produced by transfected Hep G2 cells causes hepatitis in chimpanzees. Proc Natl Acad Sci USA 84: 4641–4644PubMedCrossRefGoogle Scholar
  2. 2.
    Aldrich CE, Coates L, Wu T-T, Newbold J, Tennant BC, Summers J, Seeger C, Mason WS (1989) In vitro infection of woodchuck hepatocytes with woodchuck hepatitis virus and ground squirrel hepatitis virus. Virology 172: 247–252PubMedCrossRefGoogle Scholar
  3. 3.
    Bchini R, Capel F, Dauguet C, Dubanchet S, Petit M-A (1990) In vitro infection of human hepatoma (HepG2) cells with hepatitis B virus. J Virol 64: 3025–3032PubMedGoogle Scholar
  4. 4.
    Bonino, F, Heermann, KH, Rizzetto M, Gerlich WH (1986) Hepatitis delta virus: protein composition of delta antigen and its hepatitis B virus-derived envelope. J Virol 58: 945–950PubMedGoogle Scholar
  5. 5.
    Bonino F, Hoyer B, Shih JW-K, Rizzetto M, Purcell RH, Gerin JL (1984) Delta hepatitis agent: structural and antigenic properties of the delta-associated particle. Infect Immun 43: 1000–1005PubMedGoogle Scholar
  6. 6.
    Bruss V, Ganem D (1991) The role of envelope proteins in hepatitis B virus assembly. Proc Natl Acad Sci USA 88: 1059–1063PubMedCrossRefGoogle Scholar
  7. 7.
    Chang C, Jeng K-S, Hu C-P, Lo SJ, Su T-S, Ting L-P, Chou C-K, Han S-H, Pfaff E, Salfeld J, Schaller H (1987) Production of hepatitis B virus in vitro by transient expression of cloned HBV DNA in a hepatoma cell line. EMBO J 6: 675–680PubMedGoogle Scholar
  8. 8.
    Chen S-H, Hu C-P, Chang C (1992) Hepatitis B virus replication in well differentiated mouse hepatocyte cell lines immortalized by plasmid DNA. Cancer Res 52: 1329–1335PubMedGoogle Scholar
  9. 9.
    Chen P-J, Kalpana G, Goldberg J, Mason W, Werner B, Gerin J, Taylor J (1986) Structure and replication of the genome of the hepatitis ∂ virus. Proc Natl Acad Sci USA 83: 8774–8778PubMedCrossRefGoogle Scholar
  10. 10.
    Choi S-S, Rasshofer R, Roggendorf M (1988) Propagation of woodchuck hepatitis delta virus in primary woodchuck hepatocytes. Virology 167: 451–45PubMedGoogle Scholar
  11. 11.
    Choi S-S, Rasshofer R, Roggendorf M (1989) Inhibition of hepatitis delta virus RNA replication in primary woodchuck hepatocytes. Antiviral Res 12: 213–222PubMedCrossRefGoogle Scholar
  12. 12.
    Condreay LD, Aldrich CE, Coates L, Mason WS, Wu T-T (1990) Efficient duck hepatitis B virus production by an avian liver tumor cell line. J Virol 64: 3249–3258PubMedGoogle Scholar
  13. 13.
    Diot C, Gripon P, Rissel M, Guguen-Guillouzo C (1992) Replication of hepatitis B virus in differentiated adult rat hepatocytes transfected with cloned viral DNA. J Med Virol 36: 93–100PubMedCrossRefGoogle Scholar
  14. 14.
    Galle PR, Schlicht HJ, Fischer M, Schaller H (1988) Production of infectious duck hepatitis B virus in a human hepatoma cell line. J Virol 62: 1736–1740PubMedGoogle Scholar
  15. 15.
    Galle PR, Schlicht H-J, Kuhn C, Schaller H (1989) Replication of duck hepatitis B virus in primary duck hepatocytes and its dependence on the state of differentiation of the host cell. Hepatology 10: 459–465PubMedCrossRefGoogle Scholar
  16. 16.
    Galun E, Offensperger W-B, von Weizsäcker F, Offensperger S, Wands JR, Blum HE (1992) Human non-hepatocytes support hepadnaviral replication and virion production. J Gen Virol 73: 173–178PubMedCrossRefGoogle Scholar
  17. 17.
    Ganem D, Varmus HE (1987) The molecular biology of the hepatitis B viruses. Annu Rev Biochem 56: 651–693PubMedCrossRefGoogle Scholar
  18. 18.
    Gripon P, Diot C, Thézé N, Fourel I, Loreal O, Bréchot C, Guguen-Guillouzo C (1988) Hepatitis B virus infection of adult human hepatocytes cultured in the presence of dimethyl sulfoxide. J Virol 62: 4136–4143PubMedGoogle Scholar
  19. 19.
    Hirsh R, Colgrove R, Ganem D (1988) Replication of duck hepatitis B virus in two differentiated human hepatoma cell lines after transfection with cloned viral DNA. Virology 167: 136–142CrossRefGoogle Scholar
  20. 20.
    Hirschman SZ (1984) Replication of hepatitis B virus in cell culture systems. In: Chisari FV (ed) Advances in hepatitis research. Masson Publ., New York, pp 54–61Google Scholar
  21. 21.
    Hirschman SZ, Price P, Garfinkel E, Christman J, Acs G (1980) Expression of cloned hepatitis B virus DNA in human cell cultures. Proc Natl Acad Sci USA 77: 5507–5511PubMedCrossRefGoogle Scholar
  22. 22.
    Ichimura H, Tamura I, Tsubakio T, Kurimura O, Kurimura T (1988) Influence of hepatitis delta virus superinfection on the clearance of hepatitis B virus (HBV) markers in HBV carriers in Japan. J Med Virol 26: 49–55PubMedCrossRefGoogle Scholar
  23. 23.
    Jacob JR, Eichberg JW, Lanford RE (1989) In vitro replication and expression of hepatitis B virus from chronically infected primary chimpanzee hepatocytes. Hepatology 10: 921–927PubMedCrossRefGoogle Scholar
  24. 24.
    Kuo MY-P, Chao M, Taylor J (1989) Initiation of replication of the human hepatitis delta virus genome from cloned DNA: role of delta antigen. J Virol 63: 1945–1950PubMedGoogle Scholar
  25. 25.
    Kuo MYP, Goldberg J, Coates L, Mason W, Gerin J, Taylor J (1988) Molecular cloning of hepatitis delta virus RNA from an infected woodchuck liver: sequence, structure, and applications. J Virol 62: 1855–1861PubMedGoogle Scholar
  26. 26.
    Lanford RE, Carey KD, Estlack LE, Smith GC, Hay RV (1989) Analysis of plasma protein and lipoprotein synthesis in long-term primary cultures of baboon hepatocytes maintained in serum-free medium. In Vitro Cell Dev Biol 25: 174–182PubMedCrossRefGoogle Scholar
  27. 27.
    Müller C, Bergmann KF, Gerin JL, Korba BE (1992) Production of hepatitis B virus by stably transfected monocytic cell line U-937: a model for extrahepatic hepatitis B virus replication. J Infect Dis 165: 929–933PubMedCrossRefGoogle Scholar
  28. 28.
    Neurath AR, Kent SBH, Strick N, Parker K (1986) Identification and chemical synthesis of a host cell receptor binding site on hepatitis B virus. Cell 46: 429–436PubMedCrossRefGoogle Scholar
  29. 29.
    Ochiya T, Tsurimoto T, Ueda K, Okubo K, Shiozawa M, Matsubara K (1989) An in vitro system for infection with hepatitis B virus that uses primary human fetal hepatocytes. Proc Natl Acad Sci USA 86: 1875–1879PubMedCrossRefGoogle Scholar
  30. 30.
    Petcu DJ, Aldrich CE, Coates L, Taylor JM, Mason WS (1988) Suramin inhibits in vitro infection by duck hepatitis B virus, Rous sarcoma virus, and hepatitis delta virus. Virology 167: 385–392PubMedGoogle Scholar
  31. 31.
    Petit M-A, Dubanchet S, Capel F, Voet P, Dauguet C, Hauser P (1991) HepG2 cell binding activities of different hepatitis B virus isolates: inhibitory effect of anti-HBs and anti-preS1 (21–47). Virology 180: 483–491PubMedCrossRefGoogle Scholar
  32. 32.
    Ponzetto A, Cote PJ, Poper H, Hoyer BH, London WT, Ford EC, Bonino F, Purcell RH, Gerin JL (1984) Transmission of the hepatitis B virus-associated 9 agent to the eastern woodchuck. Proc Natl Acad Sci USA 81: 2208–2212PubMedCrossRefGoogle Scholar
  33. 33.
    Pugh JC, Summers JW (1989) Infection and uptake of duck hepatitis B virus by duck hepatocytes maintained in the presence of dimethyl sulfoxide. Virology 172: 564–572PubMedCrossRefGoogle Scholar
  34. 34.
    Pugh JC, Yaginuma K, Koike K, Summers J (1988) Duck hepatitis B virus (DHBV) particles produced by transient expression of DHBV DNA in a human hepatoma cell line are infectious in vitro. J Virol 62: 3513–3516PubMedGoogle Scholar
  35. 35.
    Rizzetto M (1983) The delta agent. Hepatology 3: 729–737PubMedCrossRefGoogle Scholar
  36. 36.
    Ryu W-S, Bayer M, Taylor J (1992) Assembly of hepatitis delta virus particles. J Virol 66: 2310–2315PubMedGoogle Scholar
  37. 37.
    Seeger C, Baldwin B, Tennant BC (1989) Expression of infectious woodchuck hepatitis virus in murine and avian fibroblasts. J Virol 63: 4665–4669PubMedGoogle Scholar
  38. 38.
    Seifer M, Heermann KH, Gerlich WH (1990) Replication of hepatitis B virus in transfected nonhepatic cells. Virology 179: 300–311PubMedCrossRefGoogle Scholar
  39. 39.
    Sells MA, Chen M-L, Acs G (1987) Production of hepatitis B virus particles in Hep G2 cells transfected with cloned hepatitis B virus DNA. Proc Natl Acad Sci USA 84: 1005–1009PubMedCrossRefGoogle Scholar
  40. 40.
    Shih C, Li L-S, Roychoudhury S, Ho M-H (1989) In vitro propagation of human hepatitis B virus in a rat hepatoma cell line. Proc Natl Acad Sci USA 86: 6323–6327PubMedCrossRefGoogle Scholar
  41. 41.
    Shih C, Yu M-YW, Li LS, Shih W-K (1990) Hepatitis B virus propagated in a rat hepatoma cell line is infectious in a primate model. Virology 179: 871–873PubMedCrossRefGoogle Scholar
  42. 42.
    Sureau C, Eichberg JW, Hubbard GB, Romet-Lemonne JL, Essex M (1988) A molecularly cloned hepatitis B virus produced in vitro is infectious in a chimpanzee. J Virol 62: 3064–3067PubMedGoogle Scholar
  43. 43.
    Sureau C, Guerra B, Lanford RE (1993) Role of the large hepatitis B virus envelope protein in infectivity of the hepatitis delta virion. J Virol 67: 366–372PubMedGoogle Scholar
  44. 44.
    Sureau C, Jacob JR, Eichberg JW, Lanford RE (1991) Tissue culture system for infection with human hepatitis delta virus. J Virol 65: 3443–3450PubMedGoogle Scholar
  45. 45.
    Sureau C, Moriarty AM, Thornton GB, Lanford RE (1992) Production of infectious hepatitis delta virus in vitro and neutralization with antibodies directed against hepatitis B virus pre-S antigens. J Virol 66: 1241–1245PubMedGoogle Scholar
  46. 46.
    Sureau C, Romet-Lemonne J-L, Mullins JI, Essex M (1986) Production of hepatitis B virus by a differentiated human hepatoma cell line after transfection with cloned circular HBV DNA. Cell 47: 37–47PubMedCrossRefGoogle Scholar
  47. 47.
    Sureau C, Taylor J, Chao M, Eichberg JW, Lanford RE (1989) Cloned hepatitis delta virus cDNA is infectious in the chimpanzee. J Virol 63: 4292–4297PubMedGoogle Scholar
  48. 48.
    Taylor J (1991) Human hepatitis delta virus. Curr Top Microbiol Immunol 168: 141–166PubMedCrossRefGoogle Scholar
  49. 49.
    Taylor J, Mason W, Summers J, Goldberg J, Aldrich C, Coates L, Gerin J, Gowans E (1987) Replication of human hepatitis delta virus in primary cultures of woodchuck hepatocytes. J Virol 61: 2891–2895PubMedGoogle Scholar
  50. 50.
    Thung, SN, Gerber, MA, Purcell RH, London WT, Mihalik KB, Popper H (1981) Animal model of human disease, chimpanzee carriers of hepatitis B virus. Am J Pathol 105: 328–332PubMedGoogle Scholar
  51. 51.
    Tiollais P, Pourcel C, Dejean A (1985) The hepatitis B virus. Nature 317: 489–495PubMedCrossRefGoogle Scholar
  52. 52.
    Tsurimoto T, Fujiyama A, Matsubara K (1987) Stable expression and replication of hepatitis B virus genome in an integrated state in a human hepatoma cell line transfected with the cloned viral DNA. Proc Natl Acad Sci USA 84: 444–448PubMedCrossRefGoogle Scholar
  53. 53.
    Tuttleman JS, Pugh JC, Summers JW (1986) In vitro experimental infection of primary duck hepatocyte cultures with duck hepatitis B virus. J Virol 58: 17–25PubMedGoogle Scholar
  54. 54.
    Ueda K, Tsurimoto T, Matsubara K (1991) Three envelope proteins of hepatitis B virus: large S, middle S and major S proteins needed for the formation of Dane particles. J Virol 65: 3521–3529PubMedGoogle Scholar
  55. 55.
    Yaginuma K, Shirakata Y, Kobayashi M, Koike K (1987) Hepatitis B virus (HBV) particles are produced in a cell culture system by transient expression of transfected HBV DNA. Proc Natl Acad Sci USA 84: 2678–2682PubMedCrossRefGoogle Scholar
  56. 56.
    Wang C-J, Chen P-J, Wu J-C, Patel D, Chen D-S (1991) Small-form hepatitis B surface antigen is sufficient to help in the assembly of hepatitis delta virus-like particles. J Virol 65: 6630–6636PubMedGoogle Scholar
  57. 57.
    Wang K-S, Choo Q-L, Weiner AJ, Ou J-H, Najarian RC, Thayer RM, Mullenbach GT, Denniston KJ, Gerin JL, Houghton M (1986) Structure, sequence and expression of the hepatitis delta (∂) viral genome. Nature 323: 508–514PubMedCrossRefGoogle Scholar
  58. 58.
    Will H, Cattaneo R, Koch H-G, Darai G, Schaller H, Schellekens H, van Eerd PMCA, Deinhardt F (1982) Cloned HBV DNA causes hepatitis in chimpanzees. Nature 299: 740–742PubMedCrossRefGoogle Scholar
  59. 59.
    Wu J-C, Chen P-J, Kuo MYP, Lee S-D, Chen D-S, Ting L-P (1991) Production of hepatitis delta virus and suppression of helper hepatitis B virus in a human hepatoma cell line. J Virol 65: 1099–1104PubMedGoogle Scholar

Copyright information

© Springer-Verlag 1993

Authors and Affiliations

  • Camille Sureau
    • 1
  1. 1.Department of Virology and ImmunologySouthwest Foundation for Biomedical ResearchSan AntonioUSA

Personalised recommendations